High-throughput analysis of protein-protein interactions in picoliter-volume droplets using fluorescence polarization

Anal Chem. 2012 Apr 17;84(8):3849-54. doi: 10.1021/ac300414g. Epub 2012 Apr 4.

Abstract

Droplet-based microfluidic systems have emerged as a powerful platform for performing high-throughput biological experimentation. In addition, fluorescence polarization has been shown to be effective in reporting a diversity of bimolecular events such as protein-protein, DNA-protein, DNA-DNA, receptor-ligand, enzyme-substrate, and protein-drug interactions. Herein, we report the use of fluorescence polarization for high-throughput protein-protein interaction analysis in a droplet-based microfluidic system. To demonstrate the efficacy of the approach, we investigate the interaction between angiogenin (ANG) and antiangiogenin antibody (anti-ANG Ab) and demonstrate the efficient extraction of dissociation constants (K(D) = 10.4 ± 3.3 nM) within short time periods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Fluorescence Polarization*
  • Humans
  • Microfluidic Analytical Techniques / instrumentation
  • Proteins / chemistry*
  • Ribonuclease, Pancreatic / chemistry

Substances

  • Antibodies, Monoclonal
  • Proteins
  • angiogenin
  • Ribonuclease, Pancreatic